希尔斯山羊草1S^(s)S和3S^(s)S染色体特异dCAPS标记的建立  

Establishment of 1S^(s)S and 3S^(s)S Chromosome⁃Specific dCAPS Markers in Aegilops searsii

作　　者：倪佳俊 韩冉[2] 徐文竞 王开 祁广 曾小雪 訾妍 李豪圣[2] 刘建军[2] 汪晓璐 刘成[2] Ni Jiajun;Han Ran;Xu Wenjing;Wang Kai;Qi Guang;Zeng Xiaoxue;Zi Yan;Li Haosheng;Liu Jianjun;Wang Xiaolu;Liu Cheng(College of Agronomy,Ludong University,Yantai 264025,China;Crop Research Institute,Shandong Academy of Agricultural Sciences/National Engineering Research Center for Wheat and Maize/Key Laboratory of Wheat Biology and Genetic Improvement in Northern Huanghuai Region,Ministry of Agriculture and Rural Affairs/Shandong Wheat Technology Innovation Center,Jinan 250100,China)

机构地区：[1]鲁东大学农学院,山东烟台264025 [2]山东省农业科学院作物研究所/小麦玉米国家工程研究中心/农业农村部黄淮北部小麦生物学与遗传育种重点实验室/山东省小麦技术创新中心,山东济南250100

出　　处：《山东农业科学》2023年第5期15-21,共7页Shandong Agricultural Sciences

基　　金：山东省重点研发计划(重大科技创新工程)项目(2021LZGC025,2021LZGC009);山东省农业科学院农业科技创新工程项目(CXGC2023A15,CXGC2022A01);山东省自然科学基金项目(ZR2021QC198);国家自然科学基金项目(31971874);泰山学者工程项目(tsqn201812123)。

摘　　要：希尔斯山羊草是小麦的近缘种,具有抗病、抗逆、抗虫等优异性状,是小麦遗传改良的珍贵基因源。当前,我们创制了一批小麦-希尔斯山羊草杂交种质。然而,由于缺乏分子标记,上述种质无法被有效筛选和鉴定。为了建立希尔斯山羊草染色体特异分子标记,本研究通过对希尔斯山羊草基因组重测序,发掘希尔斯山羊草和小麦间单核苷酸多态性(SNP),选择150个多态性位点开发dCAPS标记,结果发现,相比中国春等小麦对照,利用引物AESD23和AESD105经PCR和酶切电泳后分别能在希尔斯山羊草中获得长度不同的多态性条带。利用一套中国春-希尔斯山羊草二体附加系和端体附加系对这些多态性条带进行定位,结果发现,AESD23能在中国春-希尔斯山羊草1S^(s)二体附加系和1S^(s)S端体附加系中扩增出长度为1800 bp的多态性条带,而AESD105能在中国春-希尔斯山羊草3S^(s)二体附加系和3S^(s)S端体附加系中扩增出长度为200 bp和300 bp的多态性条带。对小麦-希尔斯山羊草杂交种质检测发现,AESD23和AESD105均能对小麦-希尔斯山羊草杂交种质进行有效筛选和鉴定。因此,上述3个多态性片段可以作为dCAPS标记用于检测含希尔斯山羊草1S^(s)S或3S^(s)S染色质的材料,为检测和追踪小麦背景中希尔斯山羊草染色质提供了新的检测手段。Aegilops searsii as a relative of wheat has excellent characters such as disease resistance,stress resistance and insect resistance,so it is a precious gene source for wheat genetic improvement.At pres⁃ent,we have created a batch of wheat⁃Ae.searsii hybrid germplasms.However,due to the lack of molecular markers,these germplasms could not be effectively screened and identified.In order to establish the chromo⁃some⁃specific molecular markers of Ae.searsii,the SNPs between Ae.searsii and wheat were discovered by re⁃sequencing the genome of Ae.searsii,and 150 polymorphic loci were selected to develop dCAPS markers in this study.The results showed that different length polymorphic bands could be obtained by PCR and enzyme⁃digestion electrophoresis using primers AESD23 and AESD105 compared with the control wheat such as Chi⁃nese Spring(CS).These polymorphic bands were mapped using a set of CS⁃Ae.searsii disomic attachment lines and telomeric attachment lines.The results showed that the polymorphic bands with length of 1800 bp could be amplified by AESD23 from the CS⁃Ae.searsii 1S^(s)disomic addition line and 1S^(s)S telosomic addition line,while AESD105 could amplify the polymorphic bands with length of 200 bp and 300 bp in the CS⁃Ae.searsii 3S^(s)disomic addition line and 3S^(s)S telosomic addition line.The detection of wheat⁃Ae.searsii hybrid germ⁃plasms found that both AESD23 and AESD105 could effectively screen and identify the hybrid germplasms of wheat⁃Ae.searsii.Therefore,the above three polymorphic fragments could be used as dCAPS markers for the detection of materials containing 1S^(s)S or 3S^(s)S chromatin of Ae.searsii.The results provided a new detection method for detecting and tracking the chromatin of Ae.searsi in wheat background.

关 键 词：小麦 希尔斯山羊草 基因组重测序 SNP分析 dCAPS标记 

分 类 号：S512.9[农业科学—作物学] Q78[生物学—分子生物学]